Issue 22, 2018

Versatile exclusion-based sample preparation platform for integrated rare cell isolation and analyte extraction

Abstract

Rare cell populations provide a patient-centric tool to monitor disease treatment, response, and resistance. However, understanding rare cells is a complex problem, which requires cell isolation/purification and downstream molecular interrogation – processes challenged by non-target populations, which vary patient-to-patient and change with disease. As such, cell isolation platforms must be amenable to a range of sample types while maintaining high efficiency and purity. The multiplexed technology for automated extraction (mTAE) is a versatile magnetic bead-based isolation platform that facilitates positive, negative, and combinatorial selection with integrated protein staining and nucleic acid isolation. mTAE is validated by isolating circulating tumor cells (CTCs) – a model rare cell population – from breast and prostate cancer patient samples. Negative selection yielded high efficiency capture of CTCs while positive selection yielded higher purity with an average of only 95 contaminant cells captured per milliliter of processed whole blood. With combinatorial selection, an overall increase in capture efficiency was observed, highlighting the potential significance of integrating multiple capture approaches on a single platform. Following capture (and staining), on platform nucleic acid extraction enabled the detection of androgen receptor-related transcripts from CTCs isolated from prostate cancer patients. The flexibility (e.g. negative, positive, combinatorial selection) and capabilities (e.g. isolation, protein staining, and nucleic acid extraction) of mTAE enable users to freely interrogate specific cell populations, a capability required to understand the potential of emerging rare cell populations and readily adapt to the heterogeneity presented across clinical samples.

Graphical abstract: Versatile exclusion-based sample preparation platform for integrated rare cell isolation and analyte extraction

Supplementary files

Article information

Article type
Paper
Submitted
03 شوال 1439
Accepted
19 محرم 1440
First published
09 صفر 1440

Lab Chip, 2018,18, 3446-3458

Author version available

Versatile exclusion-based sample preparation platform for integrated rare cell isolation and analyte extraction

H. M. Pezzi, D. J. Guckenberger, J. L. Schehr, J. Rothbauer, C. Stahlfeld, A. Singh, S. Horn, Z. D. Schultz, R. M. Bade, J. M. Sperger, S. M. Berry, J. M. Lang and D. J. Beebe, Lab Chip, 2018, 18, 3446 DOI: 10.1039/C8LC00620B

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements